U.S. patent application number 14/417703 was filed with the patent office on 2015-09-10 for nucleic acid analysis apparatus, microchip for nucleic acid analysis, and method for mounting microchip in nucleic acid analysis apparatus.
The applicant listed for this patent is Sony Corporation. Invention is credited to Masayoshi Akita, Takanori Anaguchi, Toshio Watanabe.
Application Number | 20150251184 14/417703 |
Document ID | / |
Family ID | 50027669 |
Filed Date | 2015-09-10 |
United States Patent
Application |
20150251184 |
Kind Code |
A1 |
Watanabe; Toshio ; et
al. |
September 10, 2015 |
NUCLEIC ACID ANALYSIS APPARATUS, MICROCHIP FOR NUCLEIC ACID
ANALYSIS, AND METHOD FOR MOUNTING MICROCHIP IN NUCLEIC ACID
ANALYSIS APPARATUS
Abstract
There is provided a nucleic acid analysis apparatus including a
heating unit configured to apply heat by contacting a microchip,
and a chip holding unit configured to change a position between a
first holding position that holds the microchip in midair and a
second holding position that holds the microchip in contact with
the heating unit.
Inventors: |
Watanabe; Toshio; (Kanagawa,
JP) ; Akita; Masayoshi; (Tokyo, JP) ;
Anaguchi; Takanori; (Kanagawa, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Sony Corporation |
Tokyo |
|
JP |
|
|
Family ID: |
50027669 |
Appl. No.: |
14/417703 |
Filed: |
May 27, 2013 |
PCT Filed: |
May 27, 2013 |
PCT NO: |
PCT/JP2013/064647 |
371 Date: |
January 27, 2015 |
Current U.S.
Class: |
435/287.2 ;
29/559; 435/303.1 |
Current CPC
Class: |
B01L 2300/0654 20130101;
C12Q 1/6846 20130101; G01N 2035/00158 20130101; B01L 2300/0663
20130101; B01L 2300/1805 20130101; G01N 2035/00316 20130101; B01L
7/52 20130101; B01L 2300/123 20130101; B01L 2300/1833 20130101;
G01N 2035/1034 20130101; B01L 2200/04 20130101; G01N 21/6452
20130101; B01L 2200/14 20130101; B01L 2300/0816 20130101; G01N
2035/00376 20130101; B01L 7/5255 20130101; Y10T 29/49998 20150115;
B01L 2200/12 20130101; G01N 35/00029 20130101; B01L 2300/1822
20130101; B01L 2300/0609 20130101; G01N 2035/00306 20130101; B01L
2300/043 20130101 |
International
Class: |
B01L 7/00 20060101
B01L007/00; C12Q 1/68 20060101 C12Q001/68 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 3, 2012 |
JP |
2012-172986 |
Claims
1. A nucleic acid analysis apparatus, comprising: a heating unit
configured to apply heat by contacting a microchip; and a chip
holding unit configured to change a position between a first
holding position that holds the microchip in midair and a second
holding position that holds the microchip in contact with the
heating unit.
2. The nucleic acid analysis apparatus according to claim 1,
further comprising: an opening/closing structure configured from a
hinge, wherein the chip holding unit is connected by the hinge and
moves between the first holding position and the second holding
position in coordination with opening and closing of the
opening/closing structure.
3. The nucleic acid analysis apparatus according to claim 2,
wherein the chip holding unit moves to the first holding position
in coordination with an opening operation of the opening/closing
structure, and moves to the second holding position in coordination
with a closing operation of the opening/closing structure.
4. The nucleic acid analysis apparatus according to claim 3,
wherein an insertion opening of the microchip is included in the
chip holding unit, and wherein a shape of the insertion opening is
a perpendicular cross-section shape of the microchip in an
insertion direction.
5. The nucleic acid analysis apparatus according to claim 4,
wherein the chip holding unit includes a flexible member having a
hook-shaped tip portion extending in the insertion direction of the
microchip, and wherein, in a state in which the microchip is
inserted in the chip holding unit, the tip portion of the flexible
member fits into a groove formed on a side peripheral portion of
the microchip.
6. The nucleic acid analysis apparatus according to claim 5,
wherein, in a state in which the microchip is inserted in the chip
holding unit, the tip portion of the flexible member abuts only one
face of the groove formed in a V shape, and urges the microchip in
the insertion direction.
7. The nucleic acid analysis apparatus according to claim 6,
further comprising: an upper unit and a lower unit that are each
connected by the hinge in a manner that enables them to be opened
and closed, and each include the heating unit.
8. The nucleic acid analysis apparatus according to claim 7,
further comprising: a sensor configured to detect opening and
closing of the upper unit and the lower unit.
9. The nucleic acid analysis apparatus according to claim 8,
wherein a sensor configured to detect the inserted microchip is
provided in the chip holding unit.
10. The nucleic acid analysis apparatus according to claim 9,
wherein the lower unit includes a light source, a lens, an optical
filter, and a lower heater, and wherein the upper unit includes an
upper heater, a detection filter, a lens, and a detector.
11. The nucleic acid analysis apparatus according to claim 10,
wherein the light source is an LED array, and the detector is a
PDIC array.
12. The nucleic acid analysis apparatus according to claim 11,
further comprising: a reading device of an identifier attached to
the microchip.
13. A microchip for nucleic acid analysis that has a square shape,
and has a V-shaped groove formed on a side peripheral portion for
positioning a mounting position in the nucleic acid analysis
apparatus.
14. The microchip for nucleic acid analysis according to claim 13,
wherein a notch for defining an insertion direction into an
insertion opening provided in the nucleic acid analysis apparatus
is formed on one of four corners.
15. The microchip for nucleic acid analysis according to claim 14,
formed from a plurality of substrate layers of different sizes.
16. A method for mounting a microchip in a nucleic acid analysis
apparatus, the method comprising: a step of moving a chip holding
unit connected by a hinge to a first holding position for holding
the microchip in midair, in coordination with an opening operation
of an opening/closing structure by the hinge; a step of mounting
the microchip in the chip holding unit moved to the first holding
position; and a step of moving the chip holding unit in which the
microchip is mounted to a second holding position for holding the
microchip in contact with a heating unit, in coordination with a
closing operation of the opening/closing structure.
17. An analysis apparatus, comprising: a reaction induction unit
configured to induce a reaction in a reaction area by contacting a
microchip in which the reaction area is formed; and a chip holding
unit configured to change a position between a first holding
position that holds the microchip in midair and a second holding
position that holds the microchip in contact with the reaction
induction unit.
Description
TECHNICAL FIELD
[0001] The present technology relates to a nucleic acid analysis
apparatus, a microchip for nucleic acid analysis, and a method for
mounting a microchip in that apparatus. More specifically, the
present technology relates to a nucleic acid analysis apparatus and
the like capable of accurately performing nucleic acid analysis
using a microchip.
BACKGROUND ART
[0002] An apparatus that performs nucleic acid analysis using a
substrate (microchip) formed from glass or plastic and in which a
reaction area (wells) is arranged has been disclosed. In the
nucleic acid analysis apparatus, the microchip is heated with a
heater to cause a nucleic acid amplification reaction to occur in
the wells, which include a nucleic acid that is to be amplified and
a reagent, arranged in the microchip, and the amplified nucleic
acid is optically detected.
[0003] For example, Patent Literature 1 discloses a nucleic acid
analysis apparatus that includes temperature control means for
heating a reaction area, irradiation means for irradiating light on
the reaction area, and detection means for detecting a scattered
light amount of light from the reaction area.
[0004] Regarding the present technology, there is a technology
called a hot start method for strictly controlling the reaction
time of the nucleic acid amplification reaction. The hot start
method is a method for providing an intended amplified product in a
high yield by avoiding non-specific amplification reactions caused
by misannealing of an oligonucleotide primer. In the hot start
method, the method is started by heating a mixed solution
containing reagents other than enzymes and a nucleic acid to a
denaturation temperature of the oligonucleotide primer, and adding
enzymes only after reaching the denaturation temperature.
[0005] In Patent Literature 2, in order to obtain that same effect
as for the hot start method, a "microchip for a nucleic acid
isothermal amplification reaction in which at least a part of the
substances required for the reaction, which are present in a
reaction area acting as the reaction site for an isothermal
amplification reaction of a nucleic acid in a state coated with a
thin film that melts at a temperature that is a higher than
ordinary but lower than the reaction temperature of the reaction".
In this microchip for a nucleic acid isothermal amplification
reaction, the reaction can be started at an arbitrary timing by
heating and melting the thin film coating the substances contained
in advance in the reaction area after a sample solution containing
the remaining substances and the target nucleic acid has been
supplied to the reaction area.
CITATION LIST
Patent Literature
[0006] Patent Literature 1: JP 2012-060912A [0007] Patent
Literature 2: JP 2012-024072A
SUMMARY OF INVENTION
Technical Problem
[0008] In a nucleic acid analysis apparatus, in order to obtain
stable analysis results with good reproducibility, a heater is
pre-heated to a predetermined temperature before analysis is
started. By pre-heating the heater, analysis can be started
immediately, and the non-specific amplification reactions caused by
misannealing of the oligonucleotide primer that can occur until the
heater reaches the reaction temperature can be avoided.
[0009] On the other hand, when pre-heating the heater, the nucleic
acid amplification reaction may start immediately after the
microchip has come into contact with the heater. Consequently, the
nucleic acid amplification reaction may proceed before the
apparatus has started the analysis operation after the operation
for mounting the microchip in the apparatus has been completed and
the microchip has come into contact with the heater. If such a
deviation occurs between the start time of the nucleic acid
amplification reaction and the analysis start time, the reaction
time cannot be strictly controlled, and accurate analysis results
cannot be obtained.
[0010] Accordingly, it is an object of the present invention to
provide a nucleic acid analysis apparatus capable of controlling
the timing of a reaction starting without requiring a complex
configuration or control.
Solution to Problem
[0011] In order to solve the problem, according to an embodiment of
the present disclosure, there is provided a nucleic acid analysis
apparatus including a heating unit configured to apply heat by
contacting a microchip, and a chip holding unit configured to
change a position between a first holding position that holds the
microchip in midair and a second holding position that holds the
microchip in contact with the heating unit.
[0012] The nucleic acid analysis apparatus includes an
opening/closing structure configured from a hinge. The chip holding
unit is connected by the hinge and moves between the first holding
position and the second holding position in coordination with
opening and closing of the opening/closing structure. Specifically,
the chip holding unit moves to the first holding position in
coordination with an opening operation of the opening/closing
structure, and moves to the second holding position in coordination
with a closing operation of the opening/closing structure.
[0013] In this nucleic acid analysis apparatus, by opening the
opening/closing structure and then closing the opening/closing
structure after the microchip has been held in a chip holding unit
that is at a first position, the microchip held in the chip holding
unit can be brought into contact with the heating unit for the
first time. In other words, with this nucleic acid analysis
apparatus, the microchip mounted in the chip holding unit during
opening of the opening/closing structure can be prevented from
coming into contact with the heating unit until the opening/closing
structure is closed. Therefore, with this nucleic acid analysis
apparatus, the timing of the start of the nucleic acid reaction can
be precisely controlled by making the closing operation of the
opening/closing structure and the start of heating the microchip
match.
[0014] According to an embodiment of the present disclosure, it is
preferable for the nucleic acid analysis apparatus to have an
insertion opening of the microchip in the chip holding unit. A
shape of the insertion opening is a perpendicular cross-section
shape of the microchip in an insertion direction, preferably. The
chip holding unit includes a flexible member having a hook-shaped
tip portion extending in the insertion direction of the microchip,
preferably. In a state in which the microchip is inserted in the
chip holding unit, the tip portion of the flexible member fits into
a groove formed on a side peripheral portion of the microchip. In
addition, in a state in which the microchip is inserted in the chip
holding unit, the tip portion of the flexible member abuts only one
face of the groove formed in a V shape, and urges the microchip in
the insertion direction, preferably.
[0015] According to an embodiment of the present disclosure, the
nucleic acid analysis apparatus includes an upper unit and a lower
unit that are each connected by the hinge in a manner that enables
them to be opened and closed, and each include the heating unit.
The lower unit includes a light source, a lens, an optical filter,
and a lower heater, and the upper unit includes an upper heater, a
detection filter, a lens, and a detector. The light source is an
LED array, and the detector is a PDIC array.
[0016] It is preferable for the nucleic acid analysis apparatus to
include a sensor configured to detect opening and closing of the
upper unit and the lower unit. In addition, a sensor configured to
detect the inserted microchip is provided in the chip holding unit,
preferably.
[0017] In addition, According to an embodiment of the present
disclosure, there is also provided a method for mounting a
microchip in a nucleic acid analysis apparatus, the method
including a step of moving a chip holding unit connected by a hinge
to a first holding position for holding the microchip in midair, in
coordination with an opening operation of an opening/closing
structure by the hinge, a step of mounting the microchip in the
chip holding unit moved to the first holding position, and a step
of moving the chip holding unit in which the microchip is mounted
to a second holding position for holding the microchip in contact
with a heating unit, in coordination with a closing operation of
the opening/closing structure.
[0018] In the present technology, the term "nucleic acid
amplification reaction" includes a PCR (polymerase chain reaction)
method that involves a temperature cycle, and a variety of
isothermal amplification methods that do not involve a temperature
cycle. Examples of the isothermal amplification methods include a
LAMP (Loop-Mediated Isothermal Amplification) method, an SMAP
(SMart Amplification Process) method, a NASBA (Nucleic Acid
Sequence-Based Amplification) method, an ICAN (Isothermal and
Chimeric primer-initiated Amplification of Nucleic acids) method
(registered trademark), a TRC (transcription-reverse transcription
concerted) method, a SDA (strand displacement amplification)
method, a TMA (transcription-mediated amplification) method, a RCA
(rolling circle amplification) method and the like. In addition,
the term "nucleic acid amplification reaction" widely includes
nucleic acid amplification reactions for amplifying nucleic acids
at a varying temperatures or at a constant temperature. Further,
the term "nucleic acid amplification reaction" also includes
reactions involving quantification of amplified nucleic acid
strands such as a real time PCR (RT-PCR) method and an RT-RAMP
method.
Advantageous Effects of Invention
[0019] According to the present technology, provided is a nucleic
acid analysis apparatus capable of precisely controlling the timing
of a reaction starting without requiring a complex configuration or
control.
BRIEF DESCRIPTION OF DRAWINGS
[0020] FIG. 1 is a diagram illustrating a configuration of a
nucleic acid analysis apparatus 1a according to the present
technology.
[0021] FIG. 2 is a diagram illustrating the nucleic acid analysis
apparatus 1a in a state in which an upper unit 11 and a lower unit
12 are closed.
[0022] FIG. 3 is a diagram illustrating the nucleic acid analysis
apparatus 1a in a state in which the upper unit 11 and the lower
unit 12 are open.
[0023] FIG. 4 is a diagram illustrating a configuration of the
upper unit 11 and the lower unit 12.
[0024] FIG. 5 is a diagram illustrating the nucleic acid analysis
apparatus 1a and a microchip 2 in a state in which the upper unit
11 and the lower unit 12 are open.
[0025] FIG. 6 is a diagram illustrating a configuration of a chip
holder 13.
[0026] FIG. 7 is a diagram illustrating a configuration of an
insertion opening 134 of the chip holder 13.
[0027] FIG. 8 is a diagram illustrating a configuration of a lever
135 of the chip holder 13.
[0028] FIG. 9 is a diagram illustrating a misinsertion prevention
function of the lever 135.
[0029] FIG. 10 is a diagram illustrating a positioning function of
the lever 135.
[0030] FIG. 11 is a flowchart illustrating operation of the nucleic
acid analysis apparatus 1a.
DESCRIPTION OF EMBODIMENTS
[0031] Hereinafter, preferred embodiments of the present invention
will be described in detail with reference to the appended
drawings. Note that, in this specification and the drawings,
elements that have substantially the same function and structure
are denoted with the same reference signs, and repeated explanation
is omitted. The description will be made in the following
order.
1. Nucleic acid analysis apparatus (1) Nucleic acid analysis
apparatus (2) Upper unit and lower unit (3) Chip holder (3-1)
Flip-up mechanism (3-2) Misinsertion prevention mechanism
(4) Microchip
[0032] 2. Nucleic acid analysis apparatus operation 3. Analysis
apparatus
1. Nucleic Acid Analysis Apparatus
(1) Nucleic Acid Analysis Apparatus
[0033] FIGS. 1 to 3 are diagrams illustrating a configuration of a
nucleic acid analysis apparatus according to the present
technology.
[0034] A nucleic acid analysis apparatus 1a according to the
present technology includes an AC adapter 1b and an input/output
interface 1c (refer to FIG. 1). A compact display capable of
displaying simple information and keys capable of simple inputs are
arranged on an upper face of the body of the nucleic acid analysis
apparatus 1a. An LCD display may be used for the compact display.
Further, as the keys, a keypad (numeric keys), and a
below-described power button and eject button are provided. The
input/output interface 1c includes, for example, a printer 1c1 for
outputting nucleic acid analysis results, and a code reader 1c2 for
reading an identifier attached to a microchip for nucleic acid
analysis (hereinafter also simply referred to as "microchip") that
is mounted in the apparatus. The input/output interface 1c may also
include a display, a keyboard and the like that are used to output
analysis results and input an identifier, for example.
[0035] Examples of the identifier attached to the microchip include
an item in which a chip ID indicating a production number of the
microchip and the type of reagents contained in the wells, an
analyte ID indicating the origin of a sample and the like, are
readably recorded. For example, a commonly-used barcode may be
employed.
[0036] The nucleic acid analysis apparatus 1a includes an upper
unit 11, a lower unit 12, and a chip holder 13 (refer to FIG. 2).
The upper unit 11 and the lower unit 12 are connected in a manner
that enables them to be opened and closed. Further, the chip holder
13 is also connected by a hinge 14 to both the upper unit 11 and
the lower unit 12. FIG. 2 illustrates the nucleic acid analysis
apparatus 1a in a state in which the upper unit 11 and the lower
unit 12 are closed. FIG. 3 illustrates the nucleic acid analysis
apparatus 1a in a state in which the upper unit 11 and the lower
unit 12 are open. When the upper unit 11 and the lower unit 12 are
in an open state, the chip holder 13 is positioned between the
upper unit 11 and the lower unit 12. The microchip 2 is mounted in
the apparatus by inserting the microchip 2 into the chip holder 13
in a state in which the upper unit 11 and the lower unit 12 are
open.
(2) Upper Unit and Lower Unit
[0037] A configuration of the upper unit 11 and the lower unit 12
will be described with reference to FIG. 4. The upper unit 11 and
the lower unit 12 include a structure for optically detecting a
nucleic acid amplification reaction in a reaction area (hereinafter
referred to as "wells 21") of the microchip 2.
[0038] Examples of the detection method of the nucleic acid
amplification reaction in the nucleic acid analysis apparatus 1a
include, but are not especially limited to, a method that uses a
fluorescent reagent that produces fluorescence or is extinguished
based on the progress of the nucleic acid amplification reaction,
and a method that detect changes in light scattering or absorption
or changes in polarized light in a reaction solution that occur as
the nucleic acid amplification reaction progresses.
[0039] First, the configuration of the lower unit 12 will be
described in detail. The lower unit 12 includes a lower heater 121,
a light source 122, a lens 123, and an optical filter 124. It is
preferred that a plurality of the light sources 122 and lenses 123
are arranged to correspond to the plurality of wells 21 that are
arranged in the microchip 2. Further, it is preferred that an
opening 121a for transmitting the light emitted from each light
source 122 is provided in the lower heater 121.
[0040] The lower heater 121 heats a reaction solution in the wells
21 to the reaction temperature of the nucleic acid amplification
reaction by, in a state in which the upper unit 11 and the lower
unit 12 are closed, contacting the back face of the microchip 2
mounted in the chip holder 13 to apply heat to the wells 21. It is
noted that although illustration of the chip holder 13 is omitted
in the diagram, the microchip 2 is held by the chip holder 13 in a
state in which the microchip 2 is in contact with the lower heater
121 and the below-described upper heater 111.
[0041] For the heater 21, a heat block formed from a metal such as
aluminum or gold that is generally used in conventionally-known
nucleic acid analysis apparatuses may be employed. Further, the
heater 21 may be a ceramic heater, a Peltier heater, an
electrically-heated wire and the like. Further, a transparent
conductive membrane, such as an ITO heater, that is optically
transparent can also be used. It is noted that if the wells 21 are
transparent, the opening 121a does not need to be provided.
[0042] The light source 122 may be appropriately selected based on
the optical detection method. For the light source 122, a laser
light source, a white or monochrome light-emitting diode (LED), a
mercury lamp, a tungsten lamp and the like can be employed. A
combination of two or more of these light sources may be used.
Examples of laser light sources that can be used include, but are
not especially limited to, light sources that emit light, such as a
semiconductor laser, an argon ion (Ar) laser, a helium-neon
(He--Ne) laser, a dye laser, and a krypton (Kr) laser. It is
preferred that the light source 122 is configured as an LED array
having excellent uniformity. By using an LED array, the analysis
accuracy is increased by suppressing stray light, power consumption
and production costs are suppressed, and the size of the apparatus
can be reduced.
[0043] The light emitted from the light source 122 is transmitted
through the lens 123 and an excitation filter 124, passes through
the opening 121a of the lower heater 121, and is irradiated on the
wells 21. At this stage, if the lower heater 121 is not optically
transparent, the opening 121a functions as an aperture to prevent
the irradiation of light emitted from the light source 122 that has
crossed over from an adjacent well 21. It is noted that the
excitation filter 124 is arranged as necessary for selectively
transmitting light having a specific wavelength component among the
light emitted from the light source 122.
[0044] The upper unit 11 includes an upper heater 111, lenses 112
and 113, a detector 114, and detection filters 115 and 116. It is
preferred that a plurality of the lenses 112 and 113, and detectors
114 are arranged to correspond to the plurality of wells 21.
Further, it is preferred that an opening 111a for transmitting the
light from the wells 21 to the detector 114 is provided in the
upper heater 111.
[0045] When the upper unit 11 and the lower unit 12 are in a closed
state, the upper heater 111 is in contact with a front face of the
microchip 2 mounted in the chip holder 13. The upper heater 111,
which is similarly configured to the lower heater 121, heats the
reaction solution in the wells 21 to the reaction temperature of
the nucleic acid amplification reaction by applying heat to the
wells 21.
[0046] The light produced from the wells 21 due to the irradiation
of light from the light source 122 is transmitted through the
detection filter 115, the lens 112, the detection filter 116, and
the lens 113, hits the detector 114, and is detected. At this
stage, if the upper heater 111 is not optically transparent, the
opening 111a functions as an aperture to prevent light produced
from an adjacent well 21 that crosses over from being detected.
[0047] The detected light is, based on the optical detection method
of the nucleic acid amplification reaction, transmitted light,
scattered light from a reaction product, fluorescence produced from
a fluorescent reagent and the like. The detection filters 115 and
116 are arranged as necessary for selectively transmitting light
having a specific wavelength component to the detector 114.
[0048] For the detector 114, a photo diode (PD) array, an area
image sensor such as a CCD image sensor and a CMOS sensor, a PMT
(photomultiplier tube) and the like is used. It is preferred that
the detector 114 is a PDIC array having little noise. By using a
PDIC array, analysis accuracy can be increased, and the number of
parts can be reduced.
(3) Chip Holder
(3-1) Flip-Up Mechanism
[0049] FIG. 5 illustrates the chip holder 13 in a state in which
the upper unit 11 and the lower unit 12 are open, and the microchip
2 is inserted into the chip holder 13. In a state in which the
upper unit 11 and the lower unit 12 are open, as illustrated the
diagram, the chip holder 13 is positioned away from both the upper
unit 11 and the lower unit 12.
[0050] The chip holder 13 is urged in the opening direction of the
upper unit 11 about the hinge 14 by an elastic member such as a
helical torsion coil spring. Consequently, in coordination with the
opening action of the upper unit 11, the chip holder 13 is lifted
and flipped up in the same direction by the elastic member. It is
noted that the upper unit 11 is also urged in the opening direction
about the hinge 14 by an elastic member.
[0051] It is preferred that the opening angle of the chip holder 13
when the upper unit 11 and the lower unit 12 are in an open state
(the angle formed by the lower unit 12 with the chip holder 13) is
about half the opening angle of the upper unit 11 (the angle formed
by the lower unit 12 with the chip holder 11). This is to
facilitate the insertion operation of the microchip 2 into the chip
holder 13. However, the opening angle of the chip holder 13 is not
especially limited, as long as the chip holder 13 does not come
into contact with the upper unit 11 or the lower unit 12.
[0052] A mechanism for controlling the opening angle of the chip
holder 13 to an angle like that described above is provided in the
nucleic acid analysis apparatus 1a. Specifically, as illustrated in
the diagram, a braking member 15 that is in contact with the lower
unit 12 and the chip holder 3 is provided. A groove that a pin
provided on each of the lower unit 12 and the chip holder 13 fits
into is provided in the braking member 15. The pin on the chip
holder 13 side is slidably received by this groove. The braking
member 15 controls the opening angle of the chip holder 13 to a
predetermined angle by restricting the slide amount of the pin on
the chip holder 13 side.
[0053] The following configurations can also be employed as the
configuration for flipping up the chip holder 13 to a predetermined
opening angle in coordination with the opening operation of the
upper unit 11. For example, the chip holder 13 can also be flipped
up by, when opening the upper unit 11, bringing the edge of the
upper unit 11 that connects to the hinge 14 into contact with the
edge of the chip holder 13 that connects to the hinge 14 to apply a
force for opening the chip holder 13. In this case, the
above-described force starts to act when the edge of the upper unit
11 that connects to the hinge 14 and the edge of the chip holder 13
that connects to the hinge 14 come into contact after the opening
angle of the upper unit 11 has reached a predetermined angle (e.g.,
half the maximum opening angle).
[0054] Further, for example, a gear that is rotated by the opening
operation of the upper unit 11 can be provided near the hinge 14,
and the chip holder 13 flipped up by the power of a gear that
engages and rotates with this gear. In this case, by appropriately
setting the number of teeth of the two gears, the opening angle of
the chip holder 13 can be controlled to a desired angle.
[0055] In the nucleic acid analysis apparatus 1a, based on the
respective above-described configurations, the chip holder 13 is
flipped up to a predetermined opening angle in coordination with
the opening operation of the upper unit 11. At this flipped-up
position (a first holding position), the chip holder 13 is
separated from the upper unit 11 and the lower unit 12.
Consequently, the microchip 2 inserted into the chip holder 13 is
held in midair without contacting the upper heater 111 arranged in
the upper unit 11 and the lower heater 121 arranged in the lower
unit 12.
[0056] After the microchip 2 has been inserted into the chip holder
13, the chip holder 13 is restrained by the upper unit 11 so that
when the upper unit 11 and the lower unit 12 are closed, the chip
holder 13 is sandwiched between the upper unit 11 and the lower
unit 12 (refer to FIG. 2). Consequently, the chip holder 13 moves
to a position in which the microchip 2 is held in contact with the
upper heater 111 and the lower heater 121 (a second holding
position).
[0057] An opening/closing detection sensor 161 illustrated in FIG.
5 detects the opening and closing of the upper unit 11 and the
lower unit 12. When the upper unit 11 and the lower unit 12 are
closed, a protrusion 162 provided on the upper unit 11 contacts the
opening/closing detection sensor 161, and a close operation is
detected.
(3-2) Misinsertion Prevention Mechanism
[0058] The configuration of the chip holder 13 will now be
described in more detail with reference to FIGS. 6 to 10. The chip
holder 13, which is formed from a holder outer frame 131 and a
holder inner frame 132, includes a chip detection sensor 133 for
detecting the microchip 2 inserted from an insertion opening
134.
[0059] The insertion opening 134 is formed in a shape that matches
a perpendicular cross-section in the insertion direction of the
microchip 2 so that the back and front of the microchip 2 are not
mistakenly inserted. FIG. 7 illustrates a specific example of the
shape of the insertion opening 134 (7A) and the cross-sectional
shape of the microchip 2 (7B). In this example, the cross-sectional
shape of the microchip 2 is different on the front face side and
the back face side (length in the traverse direction in the
diagram), and the insertion opening 134 is made to match this
cross-sectional shape. The microchip 2 having such a
cross-sectional shape can be produced by laminating two substrate
layers having different sizes. It is noted that the cross-sectional
shape of the insertion opening 134 and the microchip 2 is not
limited to the example illustrated here.
[0060] FIG. 8 and FIG. 9 illustrate a configuration of the holder
outer frame 131. In these diagrams, illustration of the holder
inner frame 132 is omitted. A lever 135 having a hook-shaped tip
portion is provided on the holder outer frame 131 extending in the
insertion direction of the microchip 2. When the microchip 2 is
inserted, the tip portion of the lever 135 contacts a notch 23
formed by obliquely cutting one of the four corners of the
microchip 2 (refer to FIG. 9A). The lever 135 is flexible, and thus
accommodates further insertion of the microchip 2 by flexing due to
the force applied on the tip portion by the contact with the notch
23. Although the shape of the tip portion of the lever 135 is not
especially limited, it is preferred that the shape is a curve like
that illustrated in the diagrams.
[0061] Even when the microchip 2 is inserted in a mistaken
insertion direction, as illustrated in FIG. 9B, the corners of the
microchip 2 where the notch 23 is not formed engage with the curved
portion of the tip portion formed in the shape of a hook of the
lever 135. Consequently, the lever 135 prevents further insertion
of the microchip 2 without flexing. As a result, mistaken insertion
of the microchip 2 is prevented. To make the above-described
engagement occur more easily, it is preferred that the curve angle
of the curved portion of the lever 135 is set to 90.degree. with
respect to the corner of the microchip 2.
[0062] When the microchip 2 is inserted in the correct direction,
the tip portion of the lever 135 that has been flexed by the
contact with the notch 23 slides across a face on which a V groove
22 of the inserted microchip 2 is formed, and then fits into the V
groove 22 (refer to FIG. 8B). The tip portion of the lever 135 and
the V groove 22 are arranged in a positional relationship in which
when the microchip 2 has been inserted as far as a position where
the microchip 2 hits the chip detection sensor 133, the tip portion
of the lever 135 fits into the V groove 22. It is noted that,
although an example has been described here in which the V groove
22 is provided on each of two opposing faces of the microchip 2, it
is sufficient if the V groove 22 is provided on the one face that
the tip portion of the lever 135 slides across during insertion
into the chip holder 13.
[0063] Of the two faces forming the V groove 22, the tip portion of
the lever 135 that fits into the V groove 22 is in contact with
only the face positioned on the far side in the insertion
direction. The face of the V groove 22 that is in contact with the
tip portion of the lever 135 is indicated by the reference sign 22a
in FIG. 10, and the non-contact face is indicated by reference sign
22b. Due to the flexibility of the lever 135, the lever 135 applies
a force on the contact face 22a pressing in the direction indicated
by the arrow in the diagram. The microchip 2 is pressed by this
force toward the reference point indicated by reference sign 136 in
the diagram, and is positioned in the chip holder 13.
(4) Microchip
[0064] The microchip 2 to be mounted in the nucleic acid analysis
apparatus 1a is not especially limited, as long as a region (wells)
serving as the reaction site of the nucleic acid amplification
reaction is provided. It is preferred that in the microchip 2 the
above-described V groove 22 is formed on a side peripheral portion.
Further, it is preferred that the above-described notch 23 is
formed on one of the four corners.
[0065] The microchip 2 can be formed by laminating a substrate
layer in which the wells and the like are formed. The molding of
the substrate layer can be carried out by, for example, wet etching
or dry etching of a glass substrate layer, or by nano-imprinting,
injection molding, or cutting of a plastic substrate layer.
Further, the lamination of the substrate layer can be carried out
by a known method, such as thermal fusion bonding, anodic bonding,
bonding using an adhesive sheet, plasma activated bonding, and
ultrasonic bonding.
[0066] The material of the substrate layer may be various plastics,
such as polydimethylsiloxane (PDMS), PMMA (polymethyl methacrylate:
acrylic resin), PC (polycarbonate), PS (polystyrene), PP
(polypropylene), PE (polyethylene), and PET (polyethylene
terephthalate), and glass. It is preferred to select as the
substrate layer material a material that is transmissive to light
and that has little optical error due to having little intrinsic
fluorescence and a small wavelength dispersion.
2. Nucleic Acid Analysis Apparatus Operation
[0067] Next, operation of the nucleic acid analysis apparatus 1a
will be described together with the procedure for mounting the
microchip 2 in the nucleic acid analysis apparatus 1a. FIG. 11
illustrates an operation flowchart of the nucleic acid analysis
apparatus 1a.
[0068] When starting analysis, first, operation of the apparatus is
started by pressing a key (power button) provided on an upper face
of the body of the nucleic acid analysis apparatus 1a.
[0069] When operation starts, the nucleic acid analysis apparatus
1a displays an input ID message on a compact display provided on
the upper face of the body to prompt the user to input the chip ID
and the analyte ID. Input of the chip ID and the analyte ID is
performed with a code reader or a keyboard included on the
input/output interface 1c. Further, after operation has started,
the nucleic acid analysis apparatus 1a starts heating of the upper
heater 111 of the upper unit 11 and the lower heater 121 of the
lower unit 12 to preheat these heaters to the reaction temperature
of the nucleic acid amplification reaction.
[0070] When ID input by the user has been confirmed, the nucleic
acid analysis apparatus 1a displays a mount chip message on the
compact display provided on the upper face of the body to prompt
the user to mount the chip. Simultaneously with this, the nucleic
acid analysis apparatus 1a releases the lock on the upper unit 11
and the lower unit 12 to enable the upper unit 11 to be opened. If
ID input by the user is not confirmed, the nucleic acid analysis
apparatus 1a maintains the lock on the upper unit 11 and the lower
unit 12 to prevent a microchip 2 with the wrong ID from being
mounted and analyzed.
[0071] When mounting the microchip 2 in the nucleic acid analysis
apparatus 1a, first, the chip holder 13 is moved to the first
holding position, in which the microchip 2 is held in midair, in
coordination with the opening operation of the upper unit 11 and
the lower unit 12. Specifically, the upper unit 11 is opened by
pressing a key (eject button) provided on the upper face of the
body of the nucleic acid analysis apparatus 1a (refer to FIGS. 3
and 5). When the upper unit 11 opens, the chip holder 13 flips up
in coordination with this, and separates from the upper unit 11 and
the lower unit 12 (first holding position).
[0072] Next, the microchip 2 is inserted from the insertion opening
134 of the chip holder 13 until it hits the chip detection sensor
133 (refer to FIG. 6B). The microchip 2 that has been inserted into
the chip holder 13 is held in midair without contacting the upper
heater 111 arranged in the upper unit 11 and the lower heater 121
arranged in the lower unit 12. Consequently, the microchip 2 can be
prevented from being heated by the pre-heated upper heater 111 and
lower heater 121.
[0073] Next, the chip holder 13 is moved to the second holding
position by bringing the microchip 2 into contact with the upper
heater 111 and the lower heater 121 in coordination with the
closing operation of the upper unit 11 and the lower unit 12.
Specifically, when the upper unit 11 and the lower unit 12 are
closed so as to restrain the chip holder 13 with the upper unit 11,
the chip holder 13 is sandwiched between the upper unit 11 and the
lower unit 12 (refer to FIG. 2). Consequently, the chip holder 13
moves to a position in which the microchip 2 is held in contact
with the upper heater 111 and the lower heater 121 (the second
holding position), and heating of the microchip 2 is started.
[0074] The nucleic acid analysis apparatus 1a automatically starts
analysis when the closing operation of the upper unit 11 and the
lower unit 12 is detected by the opening/closing detection sensor
161, and the microchip 2 is detected by the chip detection sensor
133. Consequently, in the nucleic acid analysis apparatus 1a,
immediately after the upper unit 11 and the lower unit 12 have
closed, the heating and the analysis of the microchip 2 by the
upper heater 111 and the lower heater 121 are simultaneously
started. Therefore, in the nucleic acid analysis apparatus 1a, the
start time of the nucleic acid amplification reaction and the start
time of analysis can be precisely matched, which enables the
reaction time to be strictly controlled and accurate analysis
results to be obtained with a high reproducibility.
[0075] After analysis has started, the nucleic acid analysis
apparatus 1a locks the upper unit 11 and the lower unit 12 so that
the upper unit 11 is not opened by mistake during analysis. It is
noted that even if ID input by the user has been confirmed,
analysis is not started as long as the close operation of the upper
unit 11 and the lower unit 12 has not been detected by the
opening/closing detection sensor. Further, even if the close
operation of the upper unit 11 and the lower unit 12 has been
detected by the opening/closing detection sensor, analysis is not
started as long as the microchip 2 is not detected by the chip
detection sensor.
[0076] When analysis is complete, the nucleic acid analysis
apparatus 1a outputs the analysis results to a printer, a display
or the like included in the input/output interface 1c. After
analysis has finished, the eject button is pressed to open the
upper unit 11, the chip holder 13 again moves to the first holding
position, and the microchip 2 is removed from the chip holder 13.
After the microchip 2 has been removed from the chip holder 13, the
upper unit 11 and the lower unit 12 are closed. At this stage,
forgetting to remove the microchip 2 can be prevented by
configuring so that the upper unit 11 and the lower unit 12 are not
locked while the microchip 2 still remains pressed into the chip
holder 13.
3. Analysis Apparatus
[0077] In the nucleic acid analysis apparatus 1a, after the
microchip 2 has been mounted in the chip holder 13, the microchip 2
is brought into contact with the upper heater 111 and the lower
heater 121 by closing the upper unit 11, and analysis is
simultaneously started. Therefore, in the nucleic acid analysis
apparatus 1a, in the nucleic acid analysis apparatus 1a, the time
at which the nucleic acid amplification reaction starts by heating
the wells 21 to the reaction temperature and the start time of
analysis can be precisely matched, which enables accurate analysis
results to be obtained with a high reproducibility.
[0078] The configuration of such a nucleic acid analysis apparatus
1a can also be applied to a microchip type analysis apparatus that
is used for analyzing reactions other than a nucleic acid
amplification reaction. Namely, the same configuration can be
widely applied in apparatuses that perform analysis by making a
material reaction equivalent to a nucleic acid amplification
reaction progress by physical or chemical induction equivalent to
heating. By employing the same configuration, in various types of
microchip type analysis apparatus, analysis can be started
simultaneously with the reaction being started in the wells by
bringing the microchip mounted in the chip holder into contact with
a reaction induction unit.
[0079] Additionally, the nucleic acid analysis apparatus according
to the present technology may also be configured as below.
(1)
[0080] A nucleic acid analysis apparatus, including:
[0081] a heating unit configured to apply heat by contacting a
microchip; and
[0082] a chip holding unit configured to change a position between
a first holding position that holds the microchip in midair and a
second holding position that holds the microchip in contact with
the heating unit.
(2)
[0083] The nucleic acid analysis apparatus according to (1),
further including:
[0084] an opening/closing structure configured from a hinge,
[0085] wherein the chip holding unit is connected by the hinge and
moves between the first holding position and the second holding
position in coordination with opening and closing of the
opening/closing structure.
(3)
[0086] The nucleic acid analysis apparatus according to (2),
wherein the chip holding unit moves to the first holding position
in coordination with an opening operation of the opening/closing
structure, and moves to the second holding position in coordination
with a closing operation of the opening/closing structure.
(4)
[0087] The nucleic acid analysis apparatus according to any one of
(1) to (3),
[0088] wherein an insertion opening of the microchip is included in
the chip holding unit, and
[0089] wherein a shape of the insertion opening is a perpendicular
cross-section shape of the microchip in an insertion direction.
(5)
[0090] The nucleic acid analysis apparatus according to (4),
[0091] wherein the chip holding unit includes a flexible member
having a hook-shaped tip portion extending in the insertion
direction of the microchip, and
[0092] wherein, in a state in which the microchip is inserted in
the chip holding unit, the tip portion of the flexible member fits
into a groove formed on a side peripheral portion of the
microchip.
(6)
[0093] The nucleic acid analysis apparatus according to (5),
wherein, in a state in which the microchip is inserted in the chip
holding unit, the tip portion of the flexible member abuts only one
face of the groove formed in a V shape, and urges the microchip in
the insertion direction.
(7)
[0094] The nucleic acid analysis apparatus according to any one of
(2) to (6), further including:
[0095] an upper unit and a lower unit that are each connected by
the hinge in a manner that enables them to be opened and closed,
and each include the heating unit.
(8)
[0096] The nucleic acid analysis apparatus according to (7),
wherein a sensor configured to detect opening and closing and a
protrusion that contacts the sensor are provided on the upper unit
and the lower unit.
(9)
[0097] The nucleic acid analysis apparatus according to any one of
(4) to (8), wherein a sensor configured to detect the inserted
microchip is provided in the chip holding unit.
(10)
[0098] The nucleic acid analysis apparatus according to any one of
(7) to (9),
[0099] wherein the lower unit includes a light source, a lens, an
optical filter, and a lower heater, and
[0100] wherein the upper unit includes an upper heater, a detection
filter, a lens, and a detector.
(11)
[0101] The nucleic acid analysis apparatus according to (10),
wherein the light source is an LED array, and the detector is a
PDIC array.
(12)
[0102] The nucleic acid analysis apparatus according to (1) to
(11), further including:
[0103] a reading device of an identifier attached to the
microchip.
[0104] Additionally, the microchip for nucleic acid analysis
according to the present technology may also be configured as
below.
(13)
[0105] A microchip for nucleic acid analysis that has a square
shape, and has a V-shaped groove formed on a side peripheral
portion for positioning a mounting position in the nucleic acid
analysis apparatus.
(14)
[0106] The microchip for nucleic acid analysis according to (13),
wherein a notch for defining an insertion direction into an
insertion opening provided in the nucleic acid analysis apparatus
is formed on one of four corners.
(15)
[0107] The microchip for nucleic acid analysis according to (13) or
(14), formed from a plurality of substrate layers of different
sizes.
REFERENCE SIGNS LIST
[0108] 1a nucleic acid analysis apparatus [0109] 1b AC adapter
[0110] 1c input/output interface [0111] 11 upper unit [0112] 111
upper heater [0113] 12 lower unit [0114] 121 lower heater [0115] 13
chip holder [0116] 131 holder outer frame [0117] 132 holder inner
frame [0118] 133 chip detection sensor [0119] 134 insertion opening
[0120] 135 lever [0121] 136 reference point [0122] 14 hinge [0123]
15 braking member [0124] 161 opening/closing detection sensor
[0125] 162 protrusion [0126] 2 microchip [0127] 21 reaction area
(wells) [0128] 22 V groove [0129] 22a contact face [0130] 22b
non-contact face [0131] 23 notch
* * * * *